Screening and Assessment of Pisatin Demethylase Activity (PDA )

Catherine C Wasmann; Jeffrey J Coleman

doi:10.1007/978-1-0716-1795-3_15

Screening and Assessment of Pisatin Demethylase Activity (PDA )

Methods Mol Biol. 2022:2391:185-190. doi: 10.1007/978-1-0716-1795-3_15.

Authors

Catherine C Wasmann¹, Jeffrey J Coleman²

Affiliations

¹ The School of Plant Sciences, University of Arizona, Tucson, AZ, USA.
² Department of Entomology and Plant Pathology, Auburn University, Auburn, AL, USA. jjcoleman@auburn.edu.

PMID: 34686986
DOI: 10.1007/978-1-0716-1795-3_15

Abstract

Plants produce low molecular weight compounds with antimicrobial activity in response to microbial attack termed phytoalexins. The first phytoalexin identified was (+) pisatin from pea, and several fungi are able to detoxify pisatin to a less inhibitory compound, including F. oxysporum f. sp. pisi. This detoxification is catalyzed by demethylation of the compound (termed pisatin demethylase activity, or PDA) by the cytochrome P450, Pda. Here we detail two procedures to assess PDA using radiolabeled [¹⁴C]pisatin as a substrate and monitoring activity using a scintillation counter.

Keywords: F. oxysporum f. sp. pisi; Host-specific virulence factor; Pea; Phytoalexin.

MeSH terms

Cytochrome P-450 Enzyme System
Oxidoreductases, O-Demethylating / metabolism*
Pterocarpans

Substances

Pterocarpans
Cytochrome P-450 Enzyme System
Oxidoreductases, O-Demethylating
pisatin demethylase
pisatin